TY - JOUR
T1 - A Genome-Scale RNAi Screen for Oct4 Modulators Defines a Role of the Paf1 Complex for Embryonic Stem Cell Identity
AU - Ding, Li
AU - Paszkowski-Rogacz, Maciej
AU - Nitzsche, Anja
AU - Slabicki, Mikolaj Michal
AU - Heninger, Anne Kristin
AU - Vries, Ingrid de
AU - Kittler, Ralf
AU - Junqueira, Magno
AU - Shevchenko, Andrej
AU - Schulz, Herbert
AU - Hubner, Norbert
AU - Doss, Michael Xavier
AU - Sachinidis, Agapios
AU - Hescheler, Juergen
AU - Iacone, Roberto
AU - Anastassiadis, Konstantinos
AU - Stewart, A. Francis
AU - Pisabarro, M. Teresa
AU - Caldarelli, Antonio
AU - Poser, Ina
AU - Theis, Mirko
AU - Buchholz, Frank
N1 - Funding Information:
We would like to thank Ina Nüsslein for help with FACS assays, Julia Jarrells for help analyzing ChIP-chip data, and Ian Chambers and Austin Smith for providing the Oct4-Gip and Sox1-GFP cell lines. This work was supported by the EU grant “FunGenES” (LSHG-CT-2003-503494), the BMBF grant GO-Bio (0315105), the Deutsche Krebshilfe grant (106685), and the DFG grant SPP1356 BU1400/3-1.
PY - 2009/5/8
Y1 - 2009/5/8
N2 - Pluripotent embryonic stem cells (ESCs) maintain self-renewal while ensuring a rapid response to differentiation cues. The identification of genes maintaining ESC identity is important to develop these cells for their potential therapeutic use. Here we report a genome-scale RNAi screen for a global survey of genes affecting ESC identity via alteration of Oct4 expression. Factors with the strongest effect on Oct4 expression included components of the Paf1 complex, a protein complex associated with RNA polymerase II. Using a combination of proteomics, expression profiling, and chromatin immunoprecipitation, we demonstrate that the Paf1C binds to promoters of key pluripotency genes, where it is required to maintain a transcriptionally active chromatin structure. The Paf1C is developmentally regulated and blocks ESC differentiation upon overexpression, and the knockdown in ESCs causes expression changes similar to Oct4 or Nanog depletions. We propose that the Paf1C plays an important role in maintaining ESC identity.
AB - Pluripotent embryonic stem cells (ESCs) maintain self-renewal while ensuring a rapid response to differentiation cues. The identification of genes maintaining ESC identity is important to develop these cells for their potential therapeutic use. Here we report a genome-scale RNAi screen for a global survey of genes affecting ESC identity via alteration of Oct4 expression. Factors with the strongest effect on Oct4 expression included components of the Paf1 complex, a protein complex associated with RNA polymerase II. Using a combination of proteomics, expression profiling, and chromatin immunoprecipitation, we demonstrate that the Paf1C binds to promoters of key pluripotency genes, where it is required to maintain a transcriptionally active chromatin structure. The Paf1C is developmentally regulated and blocks ESC differentiation upon overexpression, and the knockdown in ESCs causes expression changes similar to Oct4 or Nanog depletions. We propose that the Paf1C plays an important role in maintaining ESC identity.
KW - DNA
KW - STEMCELL
UR - http://www.scopus.com/inward/record.url?scp=65349123354&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=65349123354&partnerID=8YFLogxK
U2 - 10.1016/j.stem.2009.03.009
DO - 10.1016/j.stem.2009.03.009
M3 - Article
C2 - 19345177
AN - SCOPUS:65349123354
SN - 1934-5909
VL - 4
SP - 403
EP - 415
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 5
ER -